1. Volume 22, Issue 9, Pages 2227-2235 (February 2018)
A Single Administration of CRISPR/Cas9 Lipid Nanoparticles Achieves Robust and Persistent In Vivo Genome Editing 
Jonathan D. Finn, Amy Rhoden Smith, Mihir C. Patel, Lucinda Shaw, Madeleine R. Youniss, Jane van Heteren, Tanner Dirstine, Corey Ciullo, Reynald Lescarbeau, Jessica Seitzer, Ruchi R. Shah, Aalok Shah, Dandan Ling, Jacqueline Growe, Melissa Pink, Ellen Rohde, Kristy M. Wood, William E. Salomon, William F. Harrington, Christian Dombrowski, Walter R. Strapps, Yong Chang, David V. Morrissey 
Cell Reports 
Volume 22, Issue 9, Pages (February 2018)
DOI: /j.celrep
Copyright © 2018 Intellia Therapeutics, Inc. Terms and Conditions

2. Cell Reports 2018 22, 2227-2235DOI: (10.1016/j.celrep.2018.02.014)
Copyright © 2018 Intellia Therapeutics, Inc. Terms and Conditions

3. Figure 1
A Guide Sequence and Modification Pattern Are Critical for High Levels of In Vivo Activity
(A–F) Murine Ttr guide sequences (Table S2) were screened in vivo using a single administration of LNP-INT01 co-formulated with Cas9 mRNA and end-modified sgRNA to CD-1 mice (1 mpk of each component; 2 mpk total dose).
(A and B) 7 days post-LNP administration, mice were sacrificed (n = 5), and editing (A) and serum TTR levels were measured (B).
(C) Primary mouse hepatocytes (PMHs) were transfected with the same LNPs (10 ng/well) used in (A) and (B), and editing was measured and correlated with in vivo activity.
(D) Diagram of sgRNA with modified bases indicated in red.
(E and F) LNPs co-formulated with Cas9 mRNA and two different sgRNAs (G284, E; G269, F) targeting Ttr were administered to CD1 mice at 0.3, 1, or 2 mpk (n = 5 per group; mean and SD shown). 7 days later, liver editing was measured by NGS. Mean and SD are shown.
See also Figure S1 and Tables S1 and S2.
Cell Reports  , DOI: ( /j.celrep )
Copyright © 2018 Intellia Therapeutics, Inc. Terms and Conditions

4. Figure 2 LNP PK/PD and Long-Term Durability
(A–H) CD-1 mice were systemically administered LNPs co-formulated with Cas9 mRNA and end-modified sgRNA (G209) targeting murine Ttr.
(A and B) Mice were sacrificed (n = 3) at the indicated time points, and Cas9 mRNA and sgRNA were quantified in plasma (A) and in liver (B) (mean and SD are shown). Both Cas9 mRNA and sgRNA showed similar half-lives in plasma (T1/2 = 2.32 hr [sgRNA] vs. 2.54 hr [mRNA]) and in liver (T1/2 = 2.43 hr [sgRNA] vs. 2.09 hr [mRNA]).
(C) In a separate study, the clearance of LP01 was measured in the liver (n = 3) (dashed line = limit of quantification [LOQ]).
(D and E) CD-1 mice were administered LNP co-formulated with Cas9 mRNA and highly modified sgRNA (G211) at 2 mpk, and editing (D) and systemic TTR levels (E) were measured at the time points indicated (n = 5).
(F) CD-1 mice were administered LNP co-formulated with Cas9 mRNA and highly modified sgRNA (G211) (0.3, 1 or 3 mpk) and were sacrificed at the time points indicated (n = 5) to measure editing.
(G) Blood was collected at weeks 4, 13, and 22 from the groups sacrificed at weeks 9, 16, and 26, respectively, to measure TTR levels (mean and SEM).
(H) TTR was visualized by IHC in liver sections from mice treated with vehicle or LNPs (3 mpk) at 1 week, 6 months, or 12 months.
See also Figures S3 and S4 and Tables S1 and S2.
Cell Reports  , DOI: ( /j.celrep )
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5. Figure 3 Multi-dosing LNP-INT01 Results in Cumulative Editing
(A and B) CD-1 mice (n = 5/group) were systemically dosed with LNPs co-formulated with Cas9 mRNA and a highly modified murine Ttr-specific guide either weekly (A) or monthly (B). In the weekly dosing group, mice were dosed with 0.5 mpk on a weekly basis and were sacrificed 1 week after dosing, while the monthly group was dosed monthly and also sacrificed 1 week after dosing. As a positive control, mice were dosed with a single 2-mpk dose (black bar). Liver editing was measured by NGS, and mean + SEM are shown.
See also Tables S1 and S2.
Cell Reports  , DOI: ( /j.celrep )
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6. Figure 4 LNP-INT01 Is Effective in Rats
(A and B) Sprague Dawley rats (n = 5/group) were systemically dosed with LNPs co-formulated with Cas9 mRNA and highly modified rat-specific Ttr guides (G531, G533, and G534) at 1, 2 or 5 mpk or PBS (con). Rats were sacrificed 9 days post-administration, and liver editing (A) and systemic TTR levels were measured (B). Each data point indicates one animal with the mean and SEM indicated.
See also Tables S1 and S2.
Cell Reports  , DOI: ( /j.celrep )
Copyright © 2018 Intellia Therapeutics, Inc. Terms and Conditions